Isolation, sequencing and expression of the gene encoding a major protein from the backteriophage associated with Bartonella henselae.
The gene encoding a 31-kDa major protein (Pap31) associated with the bacteriophage harbored in Bartonella henselae was cloned and sequenced. Analysis of the resulting sequence revealed an open reading frame of 837 nucleotides coding for a protein of 279 amino acids. pap31 was then subcloned downstream of the lacZ promoter in pUC19. pap31 was amplified by polymerase chain reaction, and the linear amplicon was used as template for in-vitro transcription and translation. A protein with an apparent molecular mass of approximately 31 kDa was synthesized from this reaction. Upon analysis of the deduced aa sequence, a potential signal sequence and a consensus signal peptidase cleavage site were identified, indicative that Pap31 is modified posttranslationally, and the mature protein may be targeted to the host membrane.
Bowers
T J
TJ
University of South Florida College of Medicine, Department of Medical Microbiology and Immunology, Tampa 33612, USA.
Sweger
D
D
Jue
D
D
Anderson
B
B
D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000235:genetics* / Q000302:isolation & purification; D018416:Bartonella henselae / Q000821:virology*; D001483:Base Sequence; D003001:Cloning, Molecular; D004279:DNA, Viral; D015870:Gene Expression; D005814:Genes, Viral; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
D004279:DNA, Viral; C110491:Pap31 protein, Bartonella henselae phage 60457; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins